LOCUS       Exported                6100 bp ds-DNA     circular SYN 22-APR-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6100)
  AUTHORS   Thorsten Kaltenbacher
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 14, 2021 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6100
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..106
                     /label=scAAV-2 ITR
                     /note="5'-ITR, A' shorter than in wtAAV-2, trs "
     promoter        111..242
                     /label=hPDGF-beta promoter
                     /note="human platelet-derived growth factor beta chain 
                     promoter"
     primer_bind     238..270
                     /label=Primer 1
     primer_bind     254..289
                     /label=AAV_GibS3_1 fwd
     promoter        279..519
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        546..621
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(676..698)
                     /label=AAV_GibS3_2 rev
     primer_bind     690..708
                     /label=AAV_GibS3_3 fwd
     polyA_signal    702..909
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(902..924)
                     /label=AAV_GibS3_4 rev
     primer_bind     907..942
                     /label=AAV_GibS3_5 fwd
     polyA_site      complement(916..917)
                     /label=minimal polyA
     CDS             complement(924..1658)
                     /codon_start=1
                     /label=gfp
                     /translation="TGRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGRCHLR
                     QADPEVHLHHRQAARALAHPRDHPDLRRAVLQPLPRPHEAARLLQVRHARRLRPGAHHL
                     LQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLQQPQRLYHGRQAE
                     ERHQGELQDPPQHRGRQRAARRPLPAEHPHRRRPRVAARQPLPEHPVRPEQRPQREARS
                     HGPAGVRDRRRDHSRHGRAVQVK"
     CDS             complement(926..1645)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(1680..2301)
                     /label=RSV
     primer_bind     complement(2281..2310)
                     /label=Primer 2
     primer_bind     complement(2293..2309)
                     /label=AAV_GibS3_6 rev
     misc_feature    complement(2380..2509)
                     /label=AAV-2 ITR
                     /note="5'- and 3'-ITR, A' shorter than in wtAAV-2"
     misc_feature    2587..3801
                     /label=phage f1 (part)-1215
     rep_origin      2880..3335
                     /label=ori (f1)
                     /note="phage f1 origin of recplication"
     misc_feature    complement(3900..4174)
                     /label=pUC18-275
                     /note="fragment of pUC18;Geneious type: plasmid backbone"
     promoter        4175..4210
                     /label=AmpR promoter-36 promoter
                     /label=AmpR promoter-36
     misc_feature    4245..5105
                     /label=AmpR
                     /note="beta-lactamase, confers resistance against 
                     ampicillin"
     misc_feature    complement(5106..6051)
                     /label=pBR322-946
                     /note="part of plasmid pBR322"
     rep_origin      5260..6046
                     /label=ori (pBR322)-786
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggtaaa agcttcagtc
      121 tcctgagtag ctgggactac aggagcttgt taccacaccc agctccagtt tataaattca
      181 tctccagttt ataaaggagg aaaccgaggt actgagaggt taaaaaacct tcctgcagac
      241 acgcgtcaag cttctggatc cggtaccggc tcacatgtga gggcctattt cccatgattc
      301 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
      361 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
      421 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
      481 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggg tcttcgagaa
      541 gacctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
      601 aaagtggcac cgagtcggtg cttttttgtt ttagagctag aaatagcaag ttaaaataag
      661 gctagtccgt ttttagcgcg tgcgccaatg atcagcctcg actgtgcctt ctagttgcca
      721 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
      781 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
      841 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca
      901 tgctggggag cggccttgct agcctttact tgtacagctc gtccatgccg agagtgatcc
      961 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     1021 tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag caacacgggg ccgtcgccga
     1081 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     1141 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
     1201 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     1261 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     1321 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     1381 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
     1441 cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     1501 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     1561 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     1621 acagctcctc gcccttgctc accatggtgg cgaccggtgg atcccgggcc cgcggtaccc
     1681 agcttggagg tgcacaccaa tgtggtgaat ggtcaaatgg cgtttattgt atcgagctag
     1741 gcacttaaat acaatatctc tgcaatgcgg aattcagtgg ttcgtccaat ccatgtcaga
     1801 cccgtctgtt gccttcctaa taaggcacga tcgtaccacc ttacttccac caatcggcat
     1861 gcacggtgct ttttctctcc ttgtaaggca tgttgctaac tcatccttac catgttgcaa
     1921 gactaccaag agtatttgca taagactaca tttccccctc cctatgcaaa agcgaaacta
     1981 ctatatcctg aggggactcc taaccgcgta caaccgaagc cccgcttttc gcctaaacac
     2041 accctagtcc cctcagatac gcgtatatct ggcccgtaca tcgcgaagca gcgcaaaacg
     2101 cctaacccta agcagattct tcatgcaatt gtcggtcaag ccttgccttg ttgtagctta
     2161 aattttgctc gcgcactact cagcgacctc caacacacaa gcagggagca gatactggct
     2221 taactatgcg gcatcagagc agattgtact gagagtgcac catacggatc tgcgatgata
     2281 agctgtcaaa catgagaatt ggtcgtcgac ctgcaggcat gcaagctcta gagcatggct
     2341 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     2401 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     2461 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tggcgtaata
     2521 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     2581 gattccgttg caatggctgg cggtaatatt gttctggata ttaccagcaa ggccgatagt
     2641 ttgagttctt ctactcaggc aagtgatgtt attactaatc aaagaagtat tgcgacaacg
     2701 gttaatttgc gtgatggaca gactctttta ctcggtggcc tcactgatta taaaaacact
     2761 tctcaggatt ctggcgtacc gttcctgtct aaaatccctt taatcggcct cctgtttagc
     2821 tcccgctctg attctaacga ggaaagcacg ttatacgtgc tcgtcaaagc aaccatagta
     2881 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2941 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3001 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3061 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3121 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3181 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3241 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3301 cgcgaatttt aacaaaatat taacgcttac aatttaaata tttgcttata caatcttcct
     3361 gtttttgggg cttttctgat tatcaaccgg ggtacatatg attgacatgc tagttttacg
     3421 attaccgttc atcgattctc ttgtttgctc cagactctca ggcaatgacc tgatagcctt
     3481 tgtagagacc tctcaaaaat agctaccctc tccggcatga atttatcagc tagaacggtt
     3541 gaatatcata ttgatggtga tttgactgtc tccggccttt ctcacccgtt tgaatcttta
     3601 cctacacatt actcaggcat tgcatttaaa atatatgagg gttctaaaaa tttttatcct
     3661 tgcgttgaaa taaaggcttc tcccgcaaaa gtattacagg gtcataatgt ttttggtaca
     3721 accgatttag ctttatgctc tgaggcttta ttgcttaatt ttgctaattc tttgccttgc
     3781 ctgtatgatt tattggatgt tggaatcgcc tgatgcggta ttttctcctt acgcatctgt
     3841 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3901 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3961 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4021 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     4081 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4141 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4201 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4261 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4321 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4381 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4441 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4501 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4561 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4621 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4681 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4741 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4801 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4861 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4921 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4981 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5041 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5101 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5161 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5221 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5281 ttcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5341 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5401 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5461 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5521 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5581 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5641 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5701 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5761 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5821 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5881 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5941 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6001 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     6061 aaccgcctct ccccgcgcgt tggccgattc attaatgcag
//
